Investigation of some medicinal plants traditionally used for treatment of malaria in Kenya as potential sources of antimalarial drugs.

Malaria is a major public health problem in many tropical and subtropical countries and the burden of this disease is getting worse, mainly due to the increasing resistance of Plasmodium falciparum against the widely available antimalarial drugs. There is an urgent need for discovery of new antimalarial agents. Herbal medicines for the treatment of various diseases including malaria are an important part of the cultural diversity and traditions of which Kenya's biodiversity has been an integral part. Two major antimalarial drugs widely used today came originally from indigenous medical systems, that is quinine and artemisinin, from Peruvian and Chinese ancestral treatments, respectively. Thus ethnopharmacology is a very important resource in which new therapies may be discovered. The present review is an analysis of ethnopharmacological publications on antimalarial therapies from some Kenyan medicinal plants.

Plasmodium berghei ANKA: selection of resistance to piperaquine and lumefantrine in a mouse model.

We have selected piperaquine (PQ) and lumefantrine (LM) resistant Plasmodium berghei ANKA parasite lines in mice by drug pressure. Effective doses that reduce parasitaemia by 90% (ED(90)) of PQ and LM against the parent line were 3.52 and 3.93 mg/kg, respectively. After drug pressure (more than 27 passages), the selected parasite lines had PQ and LM resistance indexes (I(90)) [ED(90) of resistant line/ED(90) of parent line] of 68.86 and 63.55, respectively. After growing them in the absence of drug for 10 passages and cryo-preserving them at -80 degrees C for at least 2 months, the resistance phenotypes remained stable. Cross-resistance studies showed that the PQ-resistant line was highly resistant to LM, while the LM-resistant line remained sensitive to PQ. Thus, if the mechanism of resistance is similar in P. berghei and Plasmodium falciparum, the use of LM (as part of Coartem) should not select for PQ resistance.

Anti-plasmodial activity of the extracts of some Kenyan medicinal plants.

ETHNOPHARMACOLOGICAL RELEVANCE: The spread of drug resistant Plasmodium falciparum strains necessitates search for alternative newer drugs for use against malaria. Medicinal plants used traditionally in preparation of herbal medicines for malaria are potential source of new anti-malarial drugs. AIM OF THE STUDY: To identify the anti-plasmodial potential of twelve plants used in preparing herbal remedies for malaria in Kilifi and Tharaka districts of Kenya. MATERIALS AND METHODS: Twelve plants used traditionally for anti-malarial therapy in Kilifi and Tharaka districts were extracted with water/methanol yielding twenty-three extracts. The extracts were tested against chloroquine sensitive (NF54) and resistant (ENT30) P. falciparum strains in vitro using (3)Hypoxanthine assay. RESULTS: Seven (30%) extracts showed activity against P. falciparum with IC(50) values below 20 microg/ml. The remaining 16 extracts showed low or no activity. The most active extracts were from Zanthoxylum chalybeum (Rutaceae) with an IC(50) value of 3.65 microg/ml, Cyperus articulatus (Cyperaceae) with 4.84mug/ml, and Cissampelos pareira (Menispermaceae) with 5.85 microg/ml. CONCLUSIONS: This study revealed plants, that are potential sources of anti-malarial compounds. Anti-plasmodial activities of extracts of T. simplicifolia, C. pareira, and C. articulatus are reported for the first time.

The in vitro anti-plasmodial and in vivo anti-malarial efficacy of combinations of some medicinal plants used traditionally for treatment of malaria by the Meru community in Kenya.

The use of herbal drugs as combinations has existed for centuries in several cultural systems. However, the safety and efficacy of such combinations have not been validated. In this study, the toxicity, anti-plasmodial and antimalarial efficacy of several herbal drug combinations were investigated. Lannea schweinfurthii, Turraea robusta and Sclerocarya birrea, used by traditional health practitioners in Meru community, were tested for in vitro anti-plasmodial and in vivo anti-malarial activity singly against Plasmodium falciparum and Plasmodium berghei, respectively. Methanolic extract of Turraea robusta was the most active against Plasmodium falciparum D6 strain. Aqueous extracts of Lannea schweinfurthii had the highest anti-plamodial activity followed by Turraea robusta and Sclerocarya birrea. D6 was more sensitive to the plant extracts than W2 strain. Lannea schweinfurthii extracts had the highest anti-malarial activity in mice followed by Turraea robusta and Sclerocarya birrea with the methanol extracts being more active than aqueous ones. Combinations of aqueous extracts of the three plants and two others (Boscia salicifolia and Rhus natalensis) previously shown to exhibit anti-plasmodial and anti-malarial activity singly were tested in mice. Marked synergy and additive interactions were observed when combinations of the drugs were assayed in vitro. Different combinations of Turraea robusta and Lannea schweinfurthii exhibited good in vitro synergistic interactions. Combinations of Boscia salicifolia and Sclerocarya birrea; Rhus natalensis and Turraea robusta; Rhus natalensis and Boscia salicifolia; Turraea robusta and Sclerocarya birrea; and Lannea schweinfurthii and Boscia salicifolia exhibited high malaria parasite suppression (chemo-suppression >90%) in vivo when tested in mice. The findings are a preliminary demonstration of the usefulness of combining several plants in herbal drugs, as a normal practice of traditional health practitioners.

Anti-plasmodial activity of the extracts and two sesquiterpenes from Cyperus articulatus.

Two sesquiterpenes, corymbolone and mustakone, isolated from the chloroform extract of the rhizomes of Cyperus articulatus, exhibited significant anti-plasmodial properties. Mustakone was approximately ten times more active than corymbolone against the sensitive strains of the Plasmodium falciparum.

Traditional antimalarial phytotherapy remedies used by the Kwale community of the Kenyan Coast.

In Kenya, most people especially in rural areas use traditional medicine and medicinal plants to treat many diseases including malaria. Malaria is of national concern in Kenya, in view of development of resistant strains of Plasmodium falciparum to drugs especially chloroquine, which had been effective and affordable. There is need for alternative and affordable therapy. Many antimalarial drugs have been derived from medicinal plants and this is evident from the reported antiplasmodial activity. The aim of the study was to document medicinal plants traditionally used to treat malaria by the Digo community of Kwale district. Traditional health practitioners were interviewed with standardized questionnaires in order to obtain information on medicinal plants traditionally used for management of malaria. Twenty-five species in 21 genera and 16 families were encountered during the study. Celestraceae, Leguminosae and Rubiaceae families represented the species most commonly cited. Three plant species, namely; Maytenus putterlickioides, Warburgia stuhlmannii and Pentas bussei are documented for the first time for the treatment of malaria.

Antimalarial activity of some plants traditionally used in treatment of malaria in Kwale district of Kenya.

Methanolic and water extracts of five medicinal plant species used for treatment of malaria in traditional/cultural health systems of Kwale people in Kenya were tested for antimalarial activity against Plasmodium falciparum and Plasmodium berghei, respectively and for their cytotoxic effects. The most active extracts (IC(50)<10 microg/ml) screened against chloroquine (CQ) sensitive (D6) and resistant (W2) P. falciparum clones, were the water and methanol extracts of Maytenus undata (Thunb.) Blakelock (Celasteraceae), methanol extracts of Flueggea virosa (Willd.) Voigt (Euphorbiaceae), Maytenus putterlickioides (Loes.) Excell and Mendoca (Celastraceae), and Warburgia stuhlmannii Engl. (Canellaceae). These extracts showed various cytotoxic levels on Vero E6 cells with the water extract of M. undata exhibiting least cytotoxicity. At least one of the extracts of the plant species exhibited a high chemo suppression of parasitaemia >70% in a murine model of P. berghei infected mice. These results indicate that there is potential for isolation of a lead compound from the extracts of the five plants. W. stuhlmannii and M. putterlickioides have not been reported before for antiplasmodial activity.

The antiplasmodial activity of spermine alkaloids isolated from Albizia gummifera.

In the present study the methanolic extract of Albizia gummifera was fractionated into various fractions. These fractions were tested against choroquine sensitive (NF54) and resistant (ENT30) strains of Plasmodium falciparum. All other fractions apart from the alkaloidal fraction showed low activity with IC 50 above 3 microg/ml. The alkaloidal fraction exhibited strong activity against NF54 and ENT30 with IC 50 of 0.16+/-0.05 and 0.99+/-0.06 microg/ml, respectively. Five known spermine alkaloids were isolated from the alkaloidal fraction. These alkaloids exhibited activities against NF54 and ENT30 with IC 50 ranging from 0.09+/-0.02 to 0.91+/-0.10 microg/ml. Four of the alkaloids were further evaluated for in vivo activity against rodent malaria parasite Plasmodium berghei. The alkaloids showed percentage chemosuppression of parasitaemia in mice ranging from 43 to 72%. The use of the extracts A. gummifera for treatment of malaria in traditional medicine seems to have a scientific basis.

Traditional healers and the management of malaria in Kisumu district, Kenya.

OBJECTIVE: To document the ethnobotanical information on malaria treatment with the goal of eventually testing the medicinal plant extracts for antiplasmodial activity. DESIGN: A prospective study. SETTING: Informants from Kisumu City and its environs were gathered at the Kenya Medical Research Institute, Centre for Vector Biology Control Research, Kisian, Kisumu. INTERVENTIONS: Semi-structured Questionnaires were administered to 16 traditional health practitioners (THPs) to evaluate the THPs' perceptions and practice relating to causation and treatment of malaria. MAIN OUTCOME MEASURES: The THPs described the signs, symptoms and cause of malaria. Details of the preparation and use of plants for management of malaria were recorded. RESULTS: Of the 16 respondents 12 (75%) knew that malaria is transmitted by mosquito bite and 12 (75%) recognised the main symptoms as fever. Of the 36 medicinal plants, claimed to treat malaria in Kisumu, 19 plants were identified at the East African Herbarium, National Museums of Kenya. CONCLUSION: The ethnomedical and ethnobotanical data generated form the basis for pharmacological evaluation of the medicinal plants collected to establish their potential in the treatment of malaria.

Anti-plasmodial activity and toxicity of extracts of plants used in traditional malaria therapy in Meru and Kilifi Districts of Kenya.

The methanol and aqueous extracts of 10 plant species (Acacia nilotica, Azadirachta indica, Carissa edulis, Fagaropsis angolensis, Harrissonia abyssinica, Myrica salicifolia, Neoboutonia macrocalyx, Strychnos heningsii, Withania somnifera and Zanthoxylum usambarensis) used to treat malaria in Meru and Kilifi Districts, Kenya, were tested for brine shrimp lethality and in vitro anti-plasmodial activity against chloroquine-sensitive and chloroquine-resistant strains of Plasmodium falciparum (NF54 and ENT30). Of the plants tested, 40% of the methanol extracts were toxic to the brine shrimp (LD(50)<100micro/ml), while 50% showed in vitro anti-plasmodial activity (IC(50)<100microg/ml). The methanol extract of the stem bark of N. macrocalyx had the highest toxicity to brine shrimp nauplii (LD(50) 21.04+/-1.8microg/ml). Methanol extracts of the rest of the plants exhibited mild or no brine shrimp toxicity (LD(50)>50microg/ml). The aqueous extracts of N. macrocalyx had mild brine shrimp toxicity (LD(50) 41.69+/-0.9microg/ml), while the rest were lower (LD(50)>100microg/ml). The methanol extracts of F. angolensis and Zanthoxylum usambarense had IC(50) values <6microg/ml while the aqueous ones had values between 6 and 15microg/ml, against both chloroquine-sensitive and resistant P. falciparum strains. The results support the use of traditional herbs for anti-malarial therapy and demonstrate their potential as sources of drugs.

A new oleanane glycoside from the stem bark of Albizia gummifera.

The stem bark of Albizia gummifera has yielded four triterpenes, lupeol, lupenone, vitalboside-A (1) and vitalboside-A 2'-methylglucuronate (2), the last of which appears to be novel.

Triterpenes of Albizia versicolor and Albizia schimperana stem barks.

Several triterpenes were isolated from stem barks of Albizia versicolor and A. schimperana. Spectral data of acacic acid lactone (1) are presented.

Influence of manufacturing practices on quality of pharmaceutical products manufactured in Kenya.

OBJECTIVE: To establish the quality of pharmaceutical products manufactured by the respective industries in Kenya and determine the effect of manufacturing practices on the quality of these products. DESIGN: Cross-sectional study. SETTING: Industries examined are in Nairobi, Kenya. Laboratory analysis was carried out using available facilities at Kenya Medical Research Institute and University of Nairobi, Faculty of Pharmacy. INTERVENTIONS: Structured Questionnaires were administered to examine how the code of good manufacturing practices has been used in the production of each pharmaceutical product by respective companies. Questionnaires designed to evaluate the distribution and carry out limited post-market surveillance study were administered to community pharmacy outlets. Drugs were sampled and analyzed for their quality according to the respective monographs. MAIN OUTCOME MEASURES: The questionnaires administered to the industry included the source of raw materials, quarantine procedure before and after manufacture, manufacturing procedure, quality audit, quality assurance procedure, equipment, and staff. That administered to the pharmacy outlet included availability, affordability and acceptability of locally manufactured pharmaceutical products. Quality analysis of products involved the establishment of the chemical content, dissolution profile, friability, uniformity of weight and identity. For antibiotic suspensions the stability after reconstitution was also determined. RESULTS: There were 15 respondents and two non-respondents from the industry and six out of nine respondents from the pharmacy outlets. The ratio of qualified staff to product range produced seemed to influence product quality. Industries producing several products with only limited number of pharmaceutical staff had more products failing to comply with pharmacopoeia specifications compared to those producing only few products. Nevertheless, all companies are well equipped with quality control equipment, in accordance with type of product manufactured. Private pharmacies stocked few of the locally manufactured products. The reason, they said, was due to low doctor and/or patient acceptance. Compliance with quality specifications as set out in respective monographs was overall 76%. CONCLUSION: Although the local pharmaceutical industries have adopted good manufacturing practices leading to many good quality products currently in commerce, these manufacturing practices are not comprehensive and measures need to be taken to continue improving them.

Traditional herbal antimalarial therapy in Kilifi district, Kenya.

AIM OF STUDY: To identify plant species used by the traditional health practitioners (THPs) in treatment of malaria, carry out cytotoxicity and efficacy evaluation of the identified plants and to evaluate combination effects. MATERIALS AND METHODS: Thirteen plants were selected through interviews with traditional healers. In vitro antiplasmodial testing was done by measuring ability of the test sample to inhibit the incorporation of radio-labelled hypoxanthine into the malaria parasite. The extracts were tested singly and then in combination using the standard fixed ratio analysis to evaluate synergism. In vivo bioassay was done in mice using Peter's 4-days suppressive test and cytotoxicity evaluated in vitro using Vero E6 cells. RESULTS: Of the plants tested in vitro, 25% were highly active (IC(50)<10 µg/ml), 46% moderately active (IC(50) 10-50 µg/ml), 16% had weak activity of 50-100 µg/ml while 13% were not active IC(50) >100 µg/ml. Methanolic extracts of Azadirachta indica, Premna chrysoclada and Uvaria acuminata were the most active (IC(50)<10µg/ml) against both the chloroquine (CQ) sensitive (D6) and the CQ resistant (W2) Plasmodium falciparum clones. When tested in vivo in a mouse model, Azadirachta indica, Rhus natalensis and Grewia plagiophylla depicted the highest percent parasite clearance and chemo suppression of 89%, 82% and 78%, respectively. Evaluating effect of combining some of these extracts with one another against a multi-drug resistant Plasmodium falciparum (W2) clone revealed synergism among some combinations. The highest synergy was between Uvaria acuminata and Premna chrysoclada. The interaction between Grewia plagiophylla and Combretum illairii was largely antagonistic. Impressive cytotoxicity results were obtained with most of the plants tested revealing high selectivity indices an indication of enabling achievement of therapeutic doses at safe concentrations. Uvaria acuminata was, however, toxic to the cultured cells. Mild cytotoxicity was also observed in Hoslundia opposita and Lannea schweinfurthii (CC(50) 37 and 76 µg/ml, respectively). CONCLUSIONS: This study identified plants with low IC(50) values, high percent chemo suppression and low cytotoxicity thus potential sources for novel antiplasmodial agents. The findings remotely justify use of combined medicinal plants in traditional medicine practices as synergy among some plant species was demonstrated.

Antimalarial activity of some plants traditionally used in Meru district of Kenya.

Ten plant extracts commonly used by the Meru community of Kenya were evaluated for the in vitro antiplasmodial, in vivo antimalarial, cytotoxicity and animal toxicity activities. The water and methanol extracts of Ludwigia erecta and the methanol extracts of Fuerstia africana and Schkuhria pinnata exhibited high antiplasmodial activity (IC(50) < 5 microg/mL) against chloroquine sensitive (D6) and resistant (W2) Plasmodium falciparum clones. The cytotoxicity of these highly active extracts on Vero E6 cells were in the range 161.5-4650.0 microg/mL with a selectivity index (SI) of 124.2-3530.7. In vivo studies of these extracts showed less activity with chemosuppression of parasitaemia in Plasmodium berghei infected mice of 49.64-65.28%. The methanol extract of Clerodendrum eriophyllum with a lower in vitro activity (IC(50) 9.51-10.56 microg/mL) exhibited the highest chemosuppression of 90.13%. The methanol and water extracts of Pittosporum viridiflorum were toxic to mice but at a lower dose prolonged survival of P. berghei infected mice (p < 0.05) with no overt signs of toxicity. However, the extracts were cytotoxic (SI, 0.96-2.51) on Vero E6 cells. These results suggest that there is potential to isolate active non-toxic antimalarial principles from these plants.

New macrocyclic spermine (budmunchiamine) alkaloids from Albizia gummifera: with some observations on the structure--activity relationships of the budmunchiamines.

The CH2Cl2 extract of the stem bark of Albizia gummifera yielded four macrocyclic spermine alkaloids (budmunchiamines), three of them being new analogues. On the basis of spectral analysis and comparison with related compounds they were identified as budmunchiamine G (1) and the new analogues budmunchiamine K (2), 6'xi-hydroxybudmunchiamine K (3), and 9-normethylbudmunchiamine K (4). The four isolated alkaloids and other related budmunchiamines isolated from Albizia schimperana were all active against two Gram-positive and two Gram-negative bacteria at MIC levels below 80 micrograms mL(-1), and showed toxicity to brine shrimp larvae (with LC50 values below 100 micrograms mL(-1)). The negative impact of side chain hydroxylation and N-demethylation on both measures of biological activity was shown to be considerable.

Antimalarial activity of fractions isolated from Albizia gummifera and Aspilia mossambicensis crude extracts.

To identify the fractions of medicinal plant extracts containing the highest concentration of antimalarial principles, we tested the antimalarial activities of the crude or total extracts and two fractions from Albizia gummifera (Leguminosae) and three fractions from Aspilia mossambicencis (Compositae) against laboratory adapted isolates of Plasmodium falciparum in vitro using the 3H-hypoxanthine uptake assay. Chloroquine was used as a reference antimalarial drug. The mean 50% inhibitory concentrations (IC50s) of A. gummifera total extract and fraction were both <2.2 microg/ml for three P. falciparum isolates while the mean IC50 of A. gummifera f raction-2 was higher (5.0+2.9sd) for the same isolates. Aspilia mossambicensis total extract and fractions-1,4 and 5 had mean IC50 values of 96.6+/-32.5,38.6+/-23.0,142.5+/-79.6 and >1250.0 microg/ml, respectively, against four P. falciparum isolates. These results show that fraction-1 of either A. gummifera or A, mossambicensis had the highest concentration of antimalarial principles. We now plan to concentrate our efforts on these promising fractions in order to isolate pure compounds which could eventually be used to effectively treat malaria.

An in vitro evaluation of extracts from some medicinal plants in Kenya against herpes simplex virus.

The extracts from 21 medicinal plants commonly used in traditional remedies in Kenya were screened for antiviral activity against wild type 7401H strain herpes simplex virus type 1. The plant extracts exhibited antiviral activity against the virus in the plaque and yield reduction assays. The results reveal that twelve plants may contain constituents that could be exploited for the management of HSV infections. Although the extracts used in these experiments contain a complex matrix of a large number of compounds the results indicate that useful compounds can be isolated for further exploitation.